Solution Structure of the WNK1 Autoinhibitory Domain, a WNK-Specific PF2 Domain

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• 作者：Thomas M. Moon¹ ; ¹ ; ^{thomas.moon@uvm.edu} ; Fernando Correa² ; ³ ; ^{fernando.correa@utsouthwestern.edu} ; Lisa N. Kinch² ; ^{lkinch@chop.swmed.edu} ; Alexander T. Piala² ; ^{alexander.piala@utsouthwestern.edu} ; Kevin H. Gardner² ; ³ ; ^{kevin.gardner@utsouthwestern.edu} ; Elizabeth J. Goldsmith² ; ^{elizabeth.goldsmith@utsouthwestern.edu}
• 关键词：WNK1 ; with no lysine (K)-1 ; WNK1-AI ; autoinhibitory domain of WNK1 ; PF2 ; PASK/FRAY homology 2 ; HSQC ; heteronuclear single quantum coherence ; OSR ; oxidative stress responsive kinase ; SPAK ; serine/threonine proline&ndash ; alanine-rich kinase
• 刊名：Journal of Molecular Biology
• 出版年：2013
• 出版时间：26 April, 2013
• 年：2013
• 卷：425
• 期：8
• 页码：1245-1252
• 全文大小：1423 K

文摘

WNK1 [with no lysine (K)-1] is a 250-kDa serine/threonine protein kinase involved in the maintenance of cellular salt levels and is directly linked to a hereditary form of hypertension. Here, we report the solution NMR structure of the autoinhibitory domain of WNK1 (WNK1-AI), a small regulatory subunit that lies immediately C-terminal of the kinase domain. We show that this domain is a homolog of the RFXV-binding PASK/FRAY homology 2 (PF2) domain found in OSR (oxidative stress responsive) and SPAK (serine/threonine proline-alanine-rich) kinases, which are substrates of WNK1. The WNK1-AI has a circularly permuted topology relative to the OSR1-PF2 domain. Nevertheless, like PF2 domains, WNK1-AI binds peptides that contain an RFXV motif with micromolar affinities as assessed by changes in ¹H,¹⁵N heteronuclear single quantum coherence spectra. Mutations to the WNK1-AI and binding peptides confirm a similar binding mode.